TRAIL receptor associates with phosphatase SHP-1 and enables suppressing T-cell receptor signaling via inactivating Lck

We utilized whole-transcriptome RNA sequencing (RNA-seq) in TRAIL-treated activated T cells to identify the gene expression signatures and the possible regulatory pathways associated with TRAIL-R signaling on T-cell activation. CD4+ T cells were isolated from murine spleens using negative selection kits, EasySep™ Mouse CD4+ T Cell Isolation Kit (StemCell Technologies), and were cultured in RPMI-1640 medium (Invitrogen) containing 10% fetal calf serum (Invitrogen) plus penicillin (10 units/ml) and streptomycin (10 µg/ml) (Invitrogen). The CD4+ T cells were stimulated with anti-CD3 (3 µg/ml) and anti-CD28 (2 µg/ml) antibodies (BioLegend) in the presence or absence of TRAIL (10 µg/ml)(GenScript) for 24 hours. Total RNA extraction was performed using TRIzol (Invitrogen) following the manufacturer's protocol. RNA libraries for RNA-seq were prepared using Agilent SureSelect XT HS2 mRNA Library Prep Kit following manufacturer's protocols.